Technical datasheet
Description
The monoclonal antibody 8F4 recognizes mouse myeloperoxidase (MPO). MPO is a glycoprotein produced as a single precursor, which is subsequently cleaved into a alpha and beta chain. In human the biologically active MPO is a 150kDa tetramer composed of 2 glycosylated alfa chains of 59-64 kDa and 2 beta chains of 14 kDa. MPO is stored in azurophilic granules of polymorphonuclear leukocytes and is rapidly released into the phagosome and extracellular space during inflammatory conditions.The enzyme catalyzes the conversion of chloride and hydrogen peroxide to hypochlorite, a potent oxidant, which functions in host defense against microorganisms.
Involvement of MPO has been described in several human diseases, such as cardiovascular disease, airway inflammation, lung cancer, Alzheimer's disease and multiple sclerosis. A positive correlation between elevated MPO levels in serum and cardiovascular disease suggest an interesting role for MPO as an diagnostic marker, making it possible to identify patients at risk for future cardiac events. Furthermore, there are some autoimmune diseases, in which MPO is targeted by antineutrophil cytoplasm antibodies. Studies with MPO-knockout mice have shown an increased susceptibility to pneumonia following intratracheal infections. Moreover, MPO deficient mice are more susceptible to experimental autoimmune encephalitis, a T cell-dependent neuronal disease, and have an increased expression of arteriosclerotic plaques compared to wild-type mice.
The anti-mouse MPO monoclonal antibody 8F4 recognizes natural MPO in biological solutions by ELISA, in frozen tissue sections fixed with acetone and in flow cytometry using a cell permeabilization method. The antibody 8F4 is cross-reactive with rat MPO
Cross Reactivity
| Cross reactant |
Reactivity (Yes/No) |
| Rat MPO |
Yes |
Immunogen
Purified mouse MPO from WEHI-3 cells
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.
Species
Mouse IgG1
Application
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F3,4
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FC2,5
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FS
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IA1,3
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IF
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IP
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P
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W
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Yes
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No
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N.D.
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application IA has been tested by Hycult Biotech.
Use
For flow cytometry and immunohistology dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
Positive control
Neutrophils isolated from digested infarcts
Negative control
Lymphocytes isolated from digested infarcts
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.
References
1. Huugen, D et al; Aggravation of anti-myeloperoxidase antibody-induced glomerulonephritis by bacterial lipopolysaccharide: role of tumor necrosis factor-alpha. Am J Pathol 2005, 167: 47
2. Wang, D et al; C/EBPalpha directs monocytic commitment of primary myeloid progenitors. Blood 2006, 108:1223
3. Matthijsen, M et al; Myeloperoxidase is critically involved in the induction of organ damage after renal ischemia reperfusion. Am J Pathol 2007, 171: 1743
4. Leeuwen van, M et al; Accumulation of myeloperoxidase-positive neutrophils in atherosclerotic lesions in LDLR -/- mice. Arterioscler Thromb Vasc Biol 2008, 28: 84
5. Nahrendorf, M et al; An activatable MR imaging agent reports myeloperoxidase activity in healing infarcts and detects the anti-inflammatory effects of atorvastatin on ischemia-reperfusion injury non-invasively. Circulation 2008, 117: 1153
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Also available
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HM1039
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Neutrophils, Mouse, mAb NIMP-R14
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HM1051BT
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MPO, Mouse, mAb 8F4, biotinylated
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HM1051F
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MPO, Mouse, mAb 8F4, FITC
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HM3030
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MPO, Rat, mAb 2D4
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References
1. Huugen, D et al; Aggravation of anti-myeloperoxidase antibody-induced glomerulonephritis by bacterial lipopolysaccharide: role of tumor necrosis factor-alpha. Am J Pathol 2005, 167: 47
2. Wang, D et al; C/EBPalpha directs monocytic commitment of primary myeloid progenitors. Blood 2006, 108:1223
3. Matthijsen, M et al; Myeloperoxidase is critically involved in the induction of organ damage after renal ischemia reperfusion. Am J Pathol 2007, 171: 1743
4. Leeuwen van, M et al; Accumulation of myeloperoxidase-positive neutrophils in atherosclerotic lesions in LDLR -/- mice. Arterioscler Thromb Vasc Biol 2008, 28: 84
5. Nahrendorf, M et al; An activatable MR imaging agent reports myeloperoxidase activity in healing infarcts and detects the anti-inflammatory effects of atorvastatin on ischemia-reperfusion injury non-invasively. Circulation 2008, 117: 1153